[show abstract][hide abstract] ABSTRACT: The small ruminant parasite Haemonchus contortus is the most widely used parasitic nematode in drug discovery, vaccine development and anthelmintic resistance research. Its remarkable propensity to develop resistance threatens the viability of the sheep industry in many regions of the world and provides a cautionary example of the effect of mass drug administration to control parasitic nematodes. Its phylogenetic position makes it particularly well placed for comparison with the free-living nematode Caenorhabditis elegans and the most economically important parasites of livestock and humans.
Here we report the detailed analysis of a draft genome assembly and extensive transcriptomic dataset for H. contortus. This represents the first genome to be published for a strongylid nematode and the most extensive transcriptomic dataset for any parasitic nematode reported to date. We show a general pattern of conservation of genome structure and gene content between H. contortus and C. elegans, but also a dramatic expansion of important parasite gene families. We identify genes involved in parasite-specific pathways such as blood feeding, neurological function, and drug metabolism. In particular, we describe complete gene repertoires for known drug target families, providing the most comprehensive understanding yet of the action of several important anthelmintics. Also, we identify a set of genes enriched in the parasitic stages of the lifecycle and the parasite gut that provide a rich source of vaccine and drug target candidates.
The H. contortus genome and transcriptome provides an essential platform for postgenomic research in this and other important strongylid parasites.
[show abstract][hide abstract] ABSTRACT: The liver fluke, Fasciola hepatica, is common in many parts of Great Britain. To detect liver fluke infection and to assess whether fasciolicide treatment has been successful, the faecal egg count (FEC) and faecal egg count reduction test (FECRT) are widely used. Rumen fluke is also increasingly reported from Great Britain, but its species identity is yet to be determined. Liver fluke and rumen fluke eggs are morphologically similar, which may lead to erroneous diagnoses of liver fluke infection or treatment failure. As an alternative to FEC, a coproantigen ELISA (cELISA) can be used. The potential for this test to cross-react with rumen fluke species from Great Britain has not been evaluated. Rumen fluke specimens from cattle and sheep in Scotland were identified to species level using DNA sequencing of the ITS-2 region. Subsequently, rumen and liver fluke obtained from naturally co-infected sheep were subjected to immunohistochemistry using antibodies from a commercially available cELISA kit for F. hepatica. Finally, faecal samples from naturally co-infected sheep flocks were examined by FEC and cELISA. Rumen fluke from imported and home-bred cattle and sheep in Scotland belonged to the species Calicophoron daubneyi, rather than Paramphistomum cervi, the species presumed to be most common in Great Britain. Intense staining of the gastrodermis was observed in F. hepatica but cross-reactivity with C. daubneyi was not seen. Faecal samples that contained rumen fluke eggs but not liver fluke eggs were all negative by cELISA. We conclude that C. daubneyi is the most common rumen fluke of domestic ruminants in Scotland and that cELISA reduction testing may be a valuable alternative to FECRT in herds or flocks that are co-infected with liver and rumen fluke.
[show abstract][hide abstract] ABSTRACT: Sustainable control of nematode parasites in small ruminant production is a worldwide ambition. Development of anthelmintic resistance can severely impair small ruminant production. A practical approach to reduce selection pressure for anthelmintic resistance is to treat only a proportion of the flock (Targeted Selective Treatment), leaving a proportion of the nematode population untreated. The aim of this study was to compare the sustainability and efficacy of a performance-based marker, the Happy Factor™, a monitor of nutrient utilisation efficiency, with a routine whole flock anthelmintic treatment. In a commercial flock in the South West of Scotland, 183 Texel cross lambs were split into two matched but co-grazing groups: one group managed as routinely for the farm (RT group) and the other subjected to targeted selective treatment (TST group). All lambs from the RT group were drenched every 6 weeks during the grazing season, while anthelmintic administration in the TST group was restricted to animals that failed to reach pre-determined weight gain targets, based on an estimate of their efficiency of gross energy utilisation. Animal performance and parasitological data were recorded every two weeks. In the 20 week period of the study, anthelmintic treatments were reduced by approximately 50% in the TST group compared to a routine anthelmintic administration that would have been applied, whilst epg counts were always <500 throughout the study in both groups. Finally, there was no discernible difference in the mean bodyweight gain between the two groups.
[show abstract][hide abstract] ABSTRACT: A retrospective study of sheep lameness over three years revealed bacterial infection of a single joint causing severe lameness in 39 adult sheep; 5 cases were presented for veterinary examination within one week of onset of lameness. Diagnosis of a septic joint was based upon findings of severe lameness (defined as intense and extended duration) with thickening of the joint capsule. There was no significant joint effusion. All cases had been unresponsive to antibiotic therapy before referral. A penetration wound was responsible for single cases of sepsis of the stifle, shoulder and fetlock joints; the remaining 36 cases originated from bacteraemia. A potential primary septic focus was identified in six sheep at necropsy; a further six sheep had vegetative endocarditis. Radiography added little additional information except for sheep with infection of a growth plate, and neglected cases where animals had been lame for more than three months. A single treatment success was achieved with joint lavage and arthroscopy from four cases which had been lame for less than one week. All other lame sheep were euthanased for welfare reasons at presentation. Necropsy findings of unresponsive cases were characterised by pronounced synovial membrane proliferation and hyperaemia with fibrous tissue proliferation within the joint capsule and, in neglected cases, erosion of articular cartilage. Most joints contained small amounts of a pannus.
Small Ruminant Research - SMALL RUMINANT RES. 07/2012;
[show abstract][hide abstract] ABSTRACT: A field study was conducted in a sheep flock in the south east of Scotland with a history of ivermectin resistance in Teladorsagia circumcincta. The objective of the study was to compare the effects of single anthelmintic treatments in ewes before turn-out onto pasture that was contaminated with a moderate level of overwintered, ivermectin resistant, T. circumcincta infective larvae. The ewes were treated according to label directions with either a long acting injectable formulation of moxidectin (1mg/kg; affording up to 14weeks persistent action against macrocyclic lactone (ML)-susceptible T. circumcincta) or an oral formulation of moxidectin (0.2mg/kg; affording up to 5weeks persistent action against ML-susceptible T. circumcincta). The lambs were enrolled in the normal management of the farm, and received a total of three oral ivermectin treatments during the 16week study. The efficacy of both treatment strategies in controlling the periparturient rise in faecal nematode worm egg counts and subsequent pasture contamination was assessed from the faecal worm egg counts of the ewes and their lambs between lambing and weaning. Ewes that were treated with the oral formulation of moxidectin shed approximately 3.5 times more T. circumcincta eggs between lambing and weaning than ewes that were treated with the long acting formulation of moxidectin. This difference was reflected in the faecal worm egg counts of the lambs that were grazed alongside the different treatment groups of ewes. The results of the current study demonstrate persistent efficacy of the long acting formulation of moxidectin against an ivermectin resistant T. circumcincta population. The decreased pasture contamination after treatment could lead to improved lamb growth and a need for fewer anthelmintic treatments, thus potentially reducing one possible selection pressure for anthelmintic resistance. However, treatment with the long acting formulation of moxidectin would give rise to fewer susceptible nematodes being present in refugia, which could increase another possible selection pressure for anthelmintic resistance, depending on the subsequent grazing management of that pasture. The rationale for use of a persistent anthelmintic drug to control the periparturient rise in faecal ML-resistant T. circumcincta egg output of the ewes is discussed and potential differences in selection for macrocyclic lactone anthelmintic resistance using the different formulations of moxidectin are acknowledged.
[show abstract][hide abstract] ABSTRACT: Dicrocoeliosis was identified as the probable predisposing cause of weight loss and hepatogenous photosensitisation affecting half of a group of 14month-old ewe lambs on a farm on the Inner Hebridean island of Coll. Oral dosing of the ewe lambs with 15mg/kg albendazole only achieved a 79.2% reduction in Dicrocoelium dendriticum egg count, 21day post treatment. Thus, this report of dicrocoeliosis differs from the disease that occurs elsewhere, both in the severity and nature of its clinical signs, and in the response of the parasite to anthelmintic drug treatment. These differences could indicate the existence of a genetically divergent D. dendriticum population, which may have evolved within the unique and isolated biotope afforded by the machair on the island of Coll. Better understanding of these factors is a prerequisite for effective and sustainable disease control.
[show abstract][hide abstract] ABSTRACT: Various interacting factors have been identified to explain why health plans for nematode parasite control, based on conventional epidemiological knowledge and involving pharmaceutical treatments of their sheep hosts have become unsustainable. Of these, the emergence of anthelmintic resistance has had a major impact on the economics of sheep farming, necessitating fundamental managemental changes. This review focusses on the use of anthelmintic drugs for the control of gastrointestinal nematode infections in sheep, emphasising the need to develop sustainable strategies in the face of inevitable parasite evolution in response to exposure to anthelmintic drugs and other noxious stimuli, or favourable opportunities resulting from changing animal management and climatic factors.
[show abstract][hide abstract] ABSTRACT: The liver fluke, Fasciola hepatica, is a cause of significant economic losses in sheep farming. Lack of convenient and sensitive diagnostic tests in the live animal hampers the ability to monitor infection status and treatment efficacy. Use of a coproantigen ELISA and coproantigen reduction test, based on this ELISA, may address these issues but has, to date, only been evaluated in experimental challenge studies. We evaluated the coproantigen ELISA under working farm conditions in Scotland to assess its usefulness as a diagnostic test for liver fluke infection and as a diagnostic test to help determine the efficacy of flukicide treatment in sheep. First, liver fluke infection status was monitored longitudinally in a group of lambs, using monthly blood samples for biochemical assays and serum antibody ELISA and using monthly faecal samples for faecal egg count (FEC) and coproantigen ELISA. The average serum antibody ELISA titre became positive in September, two months ahead of faecal indicators of fluke infection. In contrast to results from experimental challenge studies, FEC and coproantigen ELISA became positive at the same time point. Secondly, treatment efficacy was measured in 100 ewes, from two farms, after treatment with triclabendazole (TCBZ) or closantel. Group level estimates of treatment efficacy were similar between faecal egg count reduction testing and coproantigen reduction testing at 7, 14 and 21 days post treatment. For individual animals, some inconsistencies between tests were observed. TCBZ treatment failure was noted on both farms, despite accurate weighing of animals and dosing of treatment products. We conclude that (1) coproantigen ELISA is a more convenient test than faecal egg counts and holds promise as a diagnostic tool for natural fluke infections in sheep but further evaluation of interpretation criteria may be needed; (2) the coproantigen ELISA has performed differently in the field compared with experimental challenge studies in sheep and (3) TCBZ-resistant fluke were present on both farms.
[show abstract][hide abstract] ABSTRACT: Anthelmintic drug resistance in livestock parasites is already widespread and in recent years there has been an increasing level of anthelmintic drug selection pressure applied to parasitic nematode populations in humans leading to concerns regarding the emergence of resistance. However, most parasitic nematodes, particularly those of humans, are difficult experimental subjects making mechanistic studies of drug resistance extremely difficult. The small ruminant parasitic nematode Haemonchus contortus is a more amenable model system to study many aspects of parasite biology and investigate the basic mechanisms and genetics of anthelmintic drug resistance. Here we report the successful introgression of ivermectin resistance genes from two independent ivermectin resistant strains, MHco4(WRS) and MHco10(CAVR), into the susceptible genome reference strain MHco3(ISE) using a backcrossing approach. A panel of microsatellite markers were used to monitor the procedure. We demonstrated that after four rounds of backcrossing, worms that were phenotypically resistant to ivermectin had a similar genetic background to the susceptible reference strain based on the bulk genotyping with 18 microsatellite loci and individual genotyping with a sub-panel of 9 microsatellite loci. In addition, a single marker, Hcms8a20, showed evidence of genetic linkage to an ivermectin resistance-conferring locus providing a starting point for more detailed studies of this genomic region to identify the causal mutation(s). This work presents a novel genetic approach to study anthelmintic resistance and provides a "proof-of-concept" of the use of forward genetics in an important model strongylid parasite of relevance to human hookworms. The resulting strains provide valuable resources for candidate gene studies, whole genome approaches and for further genetic analysis to identify ivermectin resistance loci.
[show abstract][hide abstract] ABSTRACT: Nematodirosis was diagnosed in the south-east of Scotland during two consecutive autumns in lambs which were grazed on the same field. The problem was unpredicted based on the knowledge of the pasture and animal management, and rudimentary understanding of the behaviour of free-living stages of Nematodirus battus in the region. Unlike the epidemiology that has been described in the south of England, whereby autumn infection of lambs is believed to arise from autumn hatching of eggs shed during the previous spring without prior chilling, it is concluded that the autumn nematodirosis in a particular sheep flock in Scotland most likely arose following prolonged survival of larvae hatched during the spring from eggs shed during the previous summer, following periods of cold exposure over the previous winter. The infective larvae survived in large numbers in a small, sheltered strip of rough grazing, where they would have been protected from harmful ultraviolet radiation and heavy rainfall, before infecting lambs during the autumn. Understanding of the evolutionary potential, nematode parasites to adapt to changing environmental conditions depends on a thorough clinical investigative approach, and is a prerequisite for future preventive management.
[show abstract][hide abstract] ABSTRACT: The effects of host age and immune suppression on abomasal parasitic infection in sheep were investigated following single experimental oral infections with MHco3 (ISE), MHco4 (WRS) and MHco10 (CAVR) strains of Haemonchus contortus in naïve 5-month-old crossbred lambs (n=1 per group) and 15-month-old Greyface sheep treated with methyl prednisolone acetate (n=2 per group) or without corticosteroid treatment (n=2 per group). Adult female H. contortus in 5-month-old lambs (n=1 per group) shed on average 6.5, 3.1 and 8.0 times more eggs than in 15-month-old sheep (n=4 per group) following infection with MHco3 (ISE), MHco4 (WRS) and MHco10 (CAVR) strains of H. contortus, respectively, over a period of 28 days following the commencement of patency. There was no obvious effect of age of sheep or corticosteroid treatment on the abomasal establishment of H. contortus or on in vitro assays for egg hatching or larval feeding at different concentrations of anthelmintics, although statistical analysis could not be performed due to the small group sizes.
The Veterinary Journal 09/2011; 189(3):296-301. · 2.42 Impact Factor
[show abstract][hide abstract] ABSTRACT: Sheep are hosts to numerous genera and species of helminth parasites, which are an important cause of production-limiting diseases. Their proper treatment and control requires knowledge of the epidemiology of these parasites in the region where the farm is located, including knowledge of the important species and their pathogenic effects, the role of immunity and resilience of the sheep, survival of L(3) on pasture under different conditions, and farm management practices. Use of anthelmintics must be combined with this knowledge to reduce risk of development of anthelmintic resistance, particularly with the control of gastrointestinal nematode parasites.
Veterinary Clinics of North America Food Animal Practice 03/2011; 27(1):139-56. · 2.40 Impact Factor
[show abstract][hide abstract] ABSTRACT: Over the past decade, definite changes have been recorded in the regional prevalence, seasonality and severity of fasciolosis in the UK, related to increased rainfall, or localised flooding, prompting debate about the deleterious effects of climate change. As a consequence, effective management of fasciolosis has become problematic in areas where fluke traditionally exists, leading to serious loss of production in sheep and cattle. Meanwhile, in eastern districts, there have been unexpected outbreaks of disease, resulting in production losses and concerns about welfare. This case report describes the economic consequences of fasciolosis in a commercial sheep flock in south-east Scotland. The diagnosis and consequences of triclabendazole resistance are discussed, in the context of developing economically sustainable control strategies.
[show abstract][hide abstract] ABSTRACT: This paper describes the nematode control strategy adopted by a Scottish hill sheep farmer. It provides an example of the limited use of anthelmintic drugs, targeted towards control of the periparturient rise in faecal nematode egg output in ewes and exploitation of grazing management, resulting in limited exposure of naive sheep to infective larvae on pasture. Resistance to benzimidazole, imidazothiazole and macrocyclic lactone anthelmintics, and to a combination of a macrocyclic lactone and imidazothiazole anthelmintic drugs was diagnosed. The targeted use of a persistent anthelmintic drug to control a periparturient rise in faecal nematode egg output in ewes may be unnecessary and selects strongly for resistance when the reservoir of anthelmintic-susceptible nematodes in refugia is small at the time of treatment. However, the use of a persistent anthelmintic drug in a selective proportion of ewes can be important and probably does not select strongly for resistance when the reservoir of anthelmintic-susceptible nematodes in refugia is large at the time of treatment. The former circumstances might arise on many Scottish hill sheep farms, whereas the latter may occur on upland and lowground farms, depending on previous grazing management, anthelmintic use and winter weather conditions. These factors must be taken into account when preparing sustainable health plans for nematode parasite control in individual sheep flocks.